Mutants that show increased sensitivity to hydrogen peroxide reveal an important role for the pentose phosphate pathway in protection of yeast against oxidative stress

被引:86
作者
Juhnke, H [1 ]
Krems, B [1 ]
Kotter, P [1 ]
Entian, KD [1 ]
机构
[1] UNIV FRANKFURT,BIOZENTRUM NIEDERURSEL,INST MIKROBIOL,D-60439 FRANKFURT,GERMANY
来源
MOLECULAR & GENERAL GENETICS | 1996年 / 252卷 / 04期
关键词
Saccharomyces cerevisiae; oxidative stress; pentose phosphate pathway; ribulose 5-phosphate epimerase;
D O I
10.1007/BF02173011
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have isolated several mutants of Saccharomyces cerevisiae that are sensitive to oxidative stress in a screen for elevated sensitivity to hydrogen peroxide. Two of the sixteen complementation groups obtained correspond to structural genes encoding enzymes of the pentose phosphate pathway. Allelism of the pos10 mutation (POS for peroxide sensitivity) to the zwf1/met1 mutants in the structural gene for glucose 6-phosphate dehydrogenase was reported previously. The second mutation. pos18, was complemented by transformation with a yeast genomic library. The open reading frame of the isolated gene encodes 238 amino acids. No detectable ribulose 5-phosphate epimerase activity was found in the pos18 mutant, suggesting that the corresponding structural gene is affected in this mutant. For that reason the gene was renamed RPE1 (for ribulose 5-phosphate epimerase). RPE1 was localized to chromosome X. The predicted protein has a molecular mass of 25 966 Daltons, a codon adaptation index (CAI) of 0.32, and an isoelectric point of 5.82. Database searches revealed 32 to 37% identity with ribulose 5-phosphate epimerases of Escherichia coli, Rhodospirillum, rubrum. Alcaligenes eutrophus and Solanum tuberosum. We have characterized RPE1 by testing enzyme activities in rpe1 deletion mutants and in strains that overexpress RPE1, and compared the hydrogen peroxide sensitivity of rpe1 mutants to that of ether mutants in the pentose phosphate pathway. Interestingly, all mutants tested (glucose 6-phosphate dehydrogenase, gluconate 6-ghosphate dehydrogenase, ribulose 5-phosphate epimerase, transketolase, transaldolase) are sensitive to hydrogen peroxide.
引用
收藏
页码:456 / 464
页数:9
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